Preparation Of Targeted PLGA-COOH Ultrasound Contrast Agents And Targeting Study In Vitro

PART I PREPARATION OF TARGETED PLGA-COOH ULTRASOUND CONTRAST AGENTS COVALENTLY BINDING WITH MONOCLONAL ANTIBODY AND TARGETING STUDY IN VITROObjective To prepare a kind of targeted hepatocellular carcinoma (HCC) high molecular polymer ultrasound contrast agents covalently binding with monoclonal antibody and investigate its affinity for HCC in vitro.Methods The high molecular polymer PLGA-COOH ultrasound contrast agents were produced by the double emulsion and the carbodiimide methods. Targeted high molecular polymer ultrasound contrast agents were prepared by combining PLGA-COOH ultrasound contrast agents with Hab18 monoclonal antibody using 1-ethyl-3-(3-dimethylaminopropyl) carbodi- imide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) mediated cross linking agents. Determine its characteristic and the ability of targeting to HCC cell line 7721 with light microscope and confocal laser scanning microscope and served non-targeted high molecular polymer ultrasound contrast agents as the control group.Results The average diameter of targeted high molecular polymer ultrasound contrast agents was(635.5±55) nm, in the targeting study in vitro, it was shown that many targeted high molecular polymer ultrasound contrast agents conjugated with HCC tightly, none conjugation in the control group.Conclusion Targeted HCC high molecular polymer ultrasound contrast agents were prepared successfully. The targeted ultrasound contrast agents can bind to HCC effectively in vitro.PART II PREPARATION OF TARGETED PLGA-COOH ULTRASOUND CONTRAST AGENTS COVALENTLY INTEGRATED WITH STREPTOAVIDIN AND TARGETING STUDY IN VITROObjective To prepare a new kind of polymer ultrasound contrast agents covalently integrated with streptoavidin (SA) and then combinded with the biotinylated antibody Hab18 to investigate the affinity for HCC in vitro.Methods The high molecular polymer ultrasound contrast agents integrated with SA were produced by double emulsion and the carbodiimide methods. Besides the usual properties, the connection between the high molecular polymer ultrasound contrast agents and SA were detected by the methods of infrared spectra and immunofluorescence. And in vitro the targeting ability of the contrast agents combinded with biotinylated antibody was contrasted with non-combined.Results Infrared spectra and immunofluorescent assay showed that SA successfully conjugated onto the surface of polymer contrast agents with activity. In vitro, it showed that many targeted high molecular polymer ultrasound contrast agents could conjugate with HCC tightly.Conclusion This high molecular polymer ultrasound contrast agent were prepared conjugated with SA successfully and it can target to HCC effectively in vitro after combined with the biotinylated antibody Hab18.